[Determination of transcription initiation site of chicken ovalbumin gene and construction of its expression vector].

To determine the core promoter of chicken ovalbumin gene and 5' upstream regions, the transcription initiation site of ovalbumin gene was confirmed by 5'RACE method, at the same time, the regulatory elements of chicken ovalbumin gene were determined by sequence analysis. To investigate the ability of regulatory elements to direct the exogenous gene expression, 1.5 kb fragment and 2.9 kb fragment were amplified by PCR method. Two fragments were subcloned to mammalian expression vector pGFP-N2 by recombinant DNA technology, the CMV promoter was cut off from pGFP-N2. Two expression vectors were constructed, one is the P2.9koval-GFP including promoter,first exon,first intron of chicken ovalbumin gene, and the other is the P1.5koval-GFP including first intron of chicken ovalbumin gene. Restriction enzyme digestion and DNA sequence analysis revealed that 5' upstream regions of ovalbumin gene were not only identical to those of the published chicken ovalbumin gene, but also were contained in the recombinant vector.